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International Journal of Innovations in Biological and Chemical Sciences, Vol. 3: 5-10, 2012
5
Rapid biogenic synthesis of silver nanoparticles using black pepper (Piper
nigrum) corn extract
Seema Garg Amity Institute of Applied Sciences, Amity University, Noida, 201 301, Uttar Pradesh, India
Abstract
Rapid biogenic synthesis of silver nanoparticles using extract of black pepper corn was attempted.
Aqueous solution of AgNO3 was treated with black pepper corn extract and can reduce most of the silver
ions into silver nanoparticles within 120 s of reaction time with the aid of microwave. The formation of
nanoparticles is reported by UV-vis spectrophotometer and complemented with characterization using
Transmission Electron Microscopy, X-ray Diffraction and Fourier Transform Infrared Spectroscopy.
Transmission Electron Microscopy revealed the presence of spherical silver nanoparticles with size
ranging of 5 to 50 nm. X-ray diffraction studies corroborated that the biosynthesized nanoparticles are
crystalline silver. Furthermore, this green biogenic approach was rapid and simple alternative to chemical
synthesis.
Key words: Piper nigrum, Silver nanoparticles, Microwave, Transmission Electron Microscopy, Transform
Infrared Spectroscopy
Introduction
In 21st
century, nanotechnology has emerged as a
rapidly growing field with numerous applications
in science and technology. Metal nanoparticles
have attracted considerable attention for their
unusual chemical and physical properties such
that they show great potential applications in
biotechnology, bioremediation of radioactive
wastes, gene delivery for treatment or
prevention of genetic disorder, catalysis, medical
imaging, particularly in medicine such as delivery
of antigen for vaccination, novel electronics,
optics, better drug delivery methods, chemical
deposition for environmental pollution cleanup
[1–12] inspired the scientists to develop
environment friendly procedures for the
synthesis of nanoparticles and to avoid use of
hazardous chemicals, which are traditionally
used. Their novel optical, thermal, chemical and
physical properties are because of higher surface
energy of nanoparticles compared to the bulk
solid and short mean free path of an electron in a
metal (10–100 nm) for many metals at room
temperature. Physical methods such as vapor
deposition, molecular beam epitaxy [13], etc.
require expensive high technology and are
neither suitable for mass production nor energy-
efficient. Although they are relatively more
affordable and suitable for mass production as
well as allowing tight control over the particle
size distribution in many ways, chemical methods
[14] are not environment friendly. Biosynthesis of
nanoparticles offers many advantages over the
corresponding physical and chemical methods.
Contrary to alternative physical and chemical
methods employing toxic chemicals which are
unacceptable for medical applications, biological
synthesis methods use bacteria [15–17], yeast
[18], fungi [19-21] or biomass of oat (Avena
sativa) and wheat (Triticum aestivum) have been
reported for irregular and rod shaped gold
nanoparticle (GNP) with size range spaning 10-30
nm [22], bengal gram bean(Cicer arietinum) was
also reported for gold nanotriangles [23].
Similarly for the synthesis of silver nanoparticle,
plants such as Capsicum annuum [24], Quercetin
[25] were used. In addition to the independent
synthesis of silver and gold nanoparticles various
plants such as sun dried biomass of
Cinnamommum camphora was used for synthesis
of triangular and spherical gold and silver
nanoparticles synthesis with size range between
55-80 nm [26], fruit extract of Amla (Emblica
officinalis) for silver and gold nanoparticles
synthesis with dimensions spanning 10-20 nm
and 15-25 nm respectively [27], leaf extract of
Aloe vera produced triangular and spherical silver
and gold nanoparticles [28], and neem
(Azadirachta indica) leaf broth was used for
*Corresponding author E-mail: [email protected]
International Journal of Innovations in Biological and Chemical Sciences, Vol. 3: 5-10, 2012
6
silver, gold and Ag–Au bimetallic Au core-Ag shell
nanoparticles[29]. Overall material and energy
consumptions in biological methods are
extremely lower, offering a low-cost green
alternative. A number of plants such as alfalfa
(Medicago sativa) produced gold nanoparticles of
4-10 nm size range [30]. These methods are very
efficient in the production of silver and gold
nanoparticles because of their simplicity,
accuracy and cost effectiveness, also from the
green chemistry perspective and their biomedical
application.
We herein report the synthesis of silver
nanoparticles using extract of black pepper corn
with the aid of microwave. Dried black pepper
corn is used medicinally for gastrointestinal
disorders [31]. In addition to these medicinal
uses, black pepper continues to be valued around
the world as important cooking spice. The
approach followed by us is cost efficient
alternative to conventional methods and
completely biogenic method of silver
nanoparticles synthesis.
MATERIALS AND METHODS
Synthesis and characterization of silver
nanoparticles
AgNO3 was purchased from Qualigens Fine
Chemicals, Mumbai, India. Black pepper corns
were purchased from a local market, washed to
remove any impurities, dried under sunlight to
completely remove the moisture and grinded in a
mixer grinder. 10 gm powder of black pepper
corn was taken and mixed with 100 ml of water
and was exposed to microwave for 180 s to
suppress the enzymes present in the solution.
The raw extract obtained was cooled and
centrifuged at 4000 rpm for 10 min then
supernatant was filtered with 11-micron mesh to
remove impurities. The resultant clear extract
was used for the synthesis of silver nanoparticles.
For reduction of Ag+
ions, 10 ml corn extract was
added to 50 ml of 10-3
M aqueous AgNO3 solution
and the solution mixture was exposed to
microwave radiation at a fixed frequency of 2.45
GHz and power of 450 watt. Periodically, aliquots
of the reaction solution were removed and
subjected to UV–vis spectroscopy measurements.
The formation of the silver nanoparticle is
monitored using UV–vis spectra by employing a
UV–visible Systronics double beam
spectrophotometer 2203 operated at a resolution
of 0.1 nm with optical path length of 10 mm. UV–
vis analysis of the reaction mixture was observed
for a period of 210 s.
The transmission electron microscopy (TEM)
image of the sample was obtained using FEI
Philips Morgagni 268D instrument operated at
100 kV. TEM samples of the aqueous suspension
of silver nanoparticles were prepared by placing a
drop of the suspension on carbon-coated copper
grids and the films on the TEM grids were
allowed to stand for 2 min, after which the extra
solution was removed using a blotting paper and
the grid was allowed to dry prior to
measurement.
The FTIR spectrum was recorded using Bruker –
FTIR Alpha instrument at a resolution of 4 cm-1
.
For XRD measurements silver nanoparticles
powdered sample was prepared by centrifuging
the synthesized silver nanoparticle solution at
10,000 rpm for 10 min. The solid residue layer
containing silver nanoparticle was redispersed in
sterile deionised water for three times to remove
unattached biological components to the surface
of nanoparticles that are not responsible for bio-
functionalization or capping. The pure residue
was dried perfectly in an oven overnight at 650C.
For the crystallinity studies, powder is used for X-
ray diffraction (XRD) study. The data was
obtained using Bruker X-ray diffractometer,
operated at 30 kV and 10 mA current with Cu
K(α) radiation of 1.54 nm wavelength.
RESULTS AND DISCUSSIONS
UV–visible absorbance studies
The addition of black pepper corn extract to silver
nitrate solution resulted in color change of the
solution from transparent to yellowish brown
after exposure to microwave radiation. The color
intensity increases with increased exposure of
reaction mixture to microwave radiation. These
color changes arise because of the excitation of
surface plasmon vibrations with the silver
nanoparticles (32). The metal ions reduction
occurs very rapidly and reduction of most of the
Ag+ was completed in 120 s. Above 120 s and up
to 150 s, the reaction process is drastically
reduced as the reaction rate changes to linear
phase. The surface plasmon resonance (SPR) of
silver nanoparticles produced a peak at 360 nm.
UV–vis absorbance of reaction mixture was taken
at 30 s interval (Fig. 1). Blank analysis was carried
out by exposing 50 ml of 10-3
M silver
nanoparticle solution to microwave for an
extended period of 210 s and validation is done
International Journal of Innovations in Biological and Chemical Sciences, Vol. 3: 5-10, 2012
7
by UV-vis study. It is observed that there is no
color change of aqueous AgNO3 solution
indicating the particles are not formed.
The microwave-assisted method is much faster as
compared to conventional studies with other
biological routes. The time required for the
conventional synthesis of silver nanoparticles
from other plants was 2-4hrs [33-34] and from
bacteria was 24-120 hrs [35-39] and thus are
rather slow.
Transmission electron microscopy
Transmission electron microscopy (TEM) was
performed for characterizing size and shape of
biosynthesized silver nanoparticles. The TEM
images of the prepared silver nanoparticles at 50
nm scale are shown in Fig. 2. It was observed that
Ag nanoparticles were circular in shape with
maximum particles in size range within 5 to 50
nm. It was also observed that silver nanoparticles
were evenly distributed in the sample.
Fourier transform infrared spectroscopy
Fig. 3 shows the FTIR spectra of biosynthesized
silver nanoparticles and carried out to identify
the possible interaction between protein and
silver nanoparticles. Results of FTIR study showed
sharp absorption peaks located at about
1631cm-1
and 3433 cm-1
. Absorptiion band at
1631cm-1
suggested the presence of amide
group, raised by the carbonyl stretch of proteins.
These results indicated that the carbonyl group of
proteins adsorbed strongly to metals, indicating
that proteins could have also formed a layer
along with the bio-organics, securing
nanoparticles. Absorption peak at 3433cm-1
are
assigned to OH stretching in alcohols and
phenolic compounds (40). The absorption peak
at 1631 cm-1
is close to that reported for native
proteins (41) which suggest that proteins are
interacting with biosynthesized nanoparticles
and also their secondary structure were not
affected during reaction with Ag+ ions or after
binding with Ag nanoparticles (42). These IR
spectroscopic studies confirmed that carbonyl
group of amino acid residues have strong binding
ability with metal suggesting the formation of
layer covering metal nanoparticles and acting as
capping agent to prevent agglomeration and
providing stability to the medium (43). These
results confirm the presence of possible proteins
acting as reducing and stabilizing agents.
X-ray diffraction measurements
Fig. 4 shows the XRD pattern of silver
nanoparticle synthesized from corn extract of
black pepper. A number of Bragg reflections with
2θ values of 38.030, 46.180, 63.43
0, and 77.18
0
correspond to the 111, 200, 220, and 311 sets of
lattice planes are observed which may be indexed
as the band for face centered cubic structures of
silver. The XRD pattern thus clearly illustrates
that the silver nanoparticles synthesized by the
present green method are crystalline in nature.
CONCLUSION
Synthesis of Silver nanoparticles using black
pepper corn extract provides an environmental
friendly, simple and efficient route. The
0
0.5
1
1.5
2
2.5
3
3.5
250 300 350 400 450 500
Wave length(nm)
Ab
so
rba
nc
e
30Sec
60Sec
90Sec
120Sec
150Sec
210Sec
Fig. 1: UV-Visible spectra of rapid biogenic
synthesis of silver nanoparticles at various
time interval
Fig. 2: TEM image of silver nanoparticles
International Journal of Innovations in Biological and Chemical Sciences, Vol. 3: 5-10, 2012
8
nanoparticles produced were of spherical in
shape with size range from 5 to 50 nm. The bio-
synthesized silver nanoparticles were
characterized using UV-Vis, XRD, TEM and FTIR
spectroscopic techniques. The synthesized silver
nanoparticles were seen to be stable due to the
presence of proteins which act as capping agent
and prevent the particle from aggregation. These
nanoparticles were crystalline in nature. From a
technological point of view, these obtained silver
nanoparticles have several advantages such as
more surface availability and can function
effectively. In this synthesis, material is
environmentally safer (plant material) compared
to chemically synthesized nanoparticles, which
often involve harmful chemicals or are
environmentally less acceptable. These
synthesized silver nanoparticles have various
applications in biomedical field, medical and
pharmaceutical applications as well as large scale
commercial production.
ACKNOWLEDGMENTS
I heartly acknowledge Dr.A.K.Chauhan,
Dr.B.Shukla, Dr. A.L .Verma and Dr. Sunita Rattan
of Amity University, Noida for their
encouragement and providing facilities for the
fulfilment of this study. I also acknowledge
Dr.Richa Krishna (AINT) for her support.
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Fig. 4 : XRD spectra of silver nanoparticles
Fig. 3: FTIR spectra of silver nanoparticles
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